The SensoLyte® 520 HIV Assay Kit uses a new FRET peptide substrate that incorporates HiLyte™ Fluor 488 (fluorophore) and QXL™ 520 (quencher) for continuous measurement of enzyme activities. In the intact FRET peptide, the fluorescence of HiLyte™ Fluor 488 is quenched by QXL™ 520. Upon cleavage of the FRET peptide by HIV protease, the fluorescence of HiLyte™ Fluor 488 is recovered and can be continuously monitored at excitation/emission = 490 nm/520 nm. With superior fluorescence quantum yield and longer emission wavelength, this HiLyte™ Fluor 488/QXL™ 520-based FRET peptide shows less interference from autofluorescence of test compounds and cellular components, thus providing better assay sensitivity. The assays are performed in a convenient 96-well or 384-well microplate format.
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